Abstract <p>In recent era, pest and pathogens of plants were killed by using a variety of chemicals which have adverse effect on both environment and human. To reduce such effects, there is strong need to replace these chemical pesticides with eco-friendly one. The current assessment is intended to address such problems and try to overcome them using seed priming technology under laboratory conditions in tomato plants. In the present study, various concentrations of vermicompost extracts (0, 20, 40, 60, 80, and 100%) were made in distilled water and tomato seeds were primed in these solutions for eight hours before being placed for germination under in vitro conditions. After 10 days of germination, various growth parameters were assessed, i.e., percent germination, root length, shoot length, root fresh weight, shoot fresh weight, root dry weight and shoot dry weight. In addition, biochemical estimation of various phytoconstituents was done using standard protocols, i.e., activity of various antioxidative enzymes, osmolytes, secondary metabolites, photosynthetic pigments, and non-enzymatic antioxidants. Moreover, FTIR, CHN and organic acid analysis were done to look into the structural modifications and level of biomolecules present in the seedlings. Results from the above experiments revealed significant enhancements in all these parameters in the concentration-dependent manner of vermicompost extract. Further, it also causes structural changes in the compounds of tomato seedlings, as indicated by Fourier Transform Infrared Spectroscopy (FTIR), CHN and High-performance liquid chromatography (HPLC). The study’s findings conclude that vermicompost extract can modulate tomato plants metabolic machinery in terms of boosting its growth and can take the place of artificial fertilizers. This is the first study to report changes in the phytochemical level and structure after being treated with vermicompost extract under in vitro conditions.</p>